ArticlePublisher preview available

Microencapsulation of Yarrowia lipolytica: cell viability and application in vitro ruminant diets

Authors:
Liliane Alves dos Santos Wanderley at Faculdade em Saúde Avançada
Liliane Alves dos Santos Wanderley
  • Faculdade em Saúde Avançada
Gean Pablo S. Aguiar at Community University of Chapecó Region - Unochapecó
Gean Pablo S. Aguiar
Jean Felipe Fossá Calisto
Jean Felipe Fossá Calisto
Jean Felipe Fossá Calisto
  • This person is not on ResearchGate, or hasn't claimed this research yet.
Jacir Dal Magro at Community University of Chapecó Region - Unochapecó
Jacir Dal Magro
Show all 11 authorsHide
Read publisher preview
Download citation
To read the full-text of this research, you can request a copy directly from the authors.

Abstract and Figures

Microencapsulation is an alternative to increase the survival capacity of microorganisms, including Yarrowia lipolytica, a widely studied yeast that produces high-value metabolites, such as lipids, aromatic compounds, biomass, lipases, and organic acids. Thus, the present study sought to investigate the effectiveness of different wall materials and the influence of the addition of salts on the microencapsulation of Y. lipolytica, evaluating yield, relationship with cell stability, ability to survive during storage, and in vitro application of ruminant diets. The spray drying process was performed via atomization, testing 11 different compositions using maltodextrin (MD), modified starch (MS) and whey protein concentrate (WPC), Y. lipolytica (Y. lipo) cells, tripolyphosphate (TPP), and sodium erythorbate (SE). The data show a reduction in the water activity value in all treatments. The highest encapsulation yield was found in treatments using MD + TPP + Y. lipo (84.0%) and WPC + TPP + Y. lipo (81.6%). Microencapsulated particles showed a survival rate ranging from 71.61 to 99.83% after 24 h. The treatments WPC + Y. lipo, WPC + SE + Y. lipo, WPC + TPP + Y. lipo, and MD + SE + Y. lipo remained stable for up to 105 days under storage conditions. The treatment WPC + SE + Y. lipo (microencapsulated yeast) was applied in the diet of ruminants due to the greater stability of cell survival. The comparison between the WPC + SE + Y. lipo treatment, wall materials, and the non-microencapsulated yeast showed that the microencapsulated yeast obtained a higher soluble fraction, degradability potential, and release of nutrients.
Figures - available from: World Journal of Microbiology and Biotechnology
This content is subject to copyright. Terms and conditions apply.
A preview of this full-text is provided by Springer Nature.
Learn more
Content available from World Journal of Microbiology and Biotechnology
This content is subject to copyright. Terms and conditions apply.
Vol.:(0123456789)
1 3
World Journal of Microbiology and Biotechnology (2023) 39:88
https://doi.org/10.1007/s11274-023-03534-2
RESEARCH
Microencapsulation ofYarrowia lipolytica: cell viability andapplication
invitro ruminant diets
LilianeAlvesdosSantosWanderley1 · GeanPabloSilvaAguiar2 · JeanFelipeFossáCalisto2 ·
JacirDalMagro2 · GabrielRossato3 · ClaitonAndréZotti3 · GuilhermedeSouzaHassemer1 ·
BrunaMariaSaorinPuton1 · RogérioLuisCansian1 · RogérioMarcosDallago1 · AlexanderJunges1
Received: 7 November 2022 / Accepted: 26 January 2023 / Published online: 6 February 2023
© The Author(s), under exclusive licence to Springer Nature B.V. 2023
Abstract
Microencapsulation is an alternative to increase the survival capacity of microorganisms, including Yarrowia lipolytica,
a widely studied yeast that produces high-value metabolites, such as lipids, aromatic compounds, biomass, lipases, and
organic acids. Thus, the present study sought to investigate the effectiveness of different wall materials and the influence of
the addition of salts on the microencapsulation of Y. lipolytica, evaluating yield, relationship with cell stability, ability to
survive during storage, and invitro application of ruminant diets. The spray drying process was performed via atomization,
testing 11 different compositions using maltodextrin (MD), modified starch (MS) and whey protein concentrate (WPC),
Y. lipolytica (Y. lipo) cells, tripolyphosphate (TPP), and sodium erythorbate (SE). The data show a reduction in the water
activity value in all treatments. The highest encapsulation yield was found in treatments using MD + TPP + Y. lipo (84.0%)
and WPC + TPP + Y. lipo (81.6%). Microencapsulated particles showed a survival rate ranging from 71.61 to 99.83% after
24h. The treatments WPC + Y. lipo, WPC + SE + Y. lipo, WPC + TPP + Y. lipo, and MD + SE + Y. lipo remained stable for
up to 105days under storage conditions. The treatment WPC + SE + Y. lipo (microencapsulated yeast) was applied in the diet
of ruminants due to the greater stability of cell survival. The comparison between the WPC + SE + Y. lipo treatment, wall
materials, and the non-microencapsulated yeast showed that the microencapsulated yeast obtained a higher soluble fraction,
degradability potential, and release of nutrients.
Keywords Microencapsulation· Yarrowia lipolytica· Spray drying· Viable cell count
* Alexander Junges
junges@uricer.edu.br
Liliane Alves dos Santos Wanderley
lilianesw@gmail.com
Gean Pablo Silva Aguiar
geanpablo@gmail.com
Jean Felipe Fossá Calisto
jeanfelipe@unochapeco.edu.br
Jacir Dal Magro
jacir@unochapeco.edu.br
Gabriel Rossato
gabrielrossato30@gmail.com
Claiton André Zotti
claiton.zotti@unoesc.edu.br
Guilherme de Souza Hassemer
guilherme.hassemer@hotmail.com
Bruna Maria Saorin Puton
brunnaputon@hotmail.com
Rogério Luis Cansian
cansian@uricer.edu.br
Rogério Marcos Dallago
dallago@uricer.edu.br
1 Department ofFood andChemical Engineering, URI
– Erechim, 1621, Sete de Setembro Av., Erechim,
RS99709-910, Brazil
2 Environmental Sciences Area, Community University
ofChapecó Region (Unochapecó), Servidão Anjo da Guarda,
295-D, Bairro Efapi, Chapecó, SC89809-900, Brazil
3 Department ofAnimal Science, University ofWest Santa
Catarina, Xanxerê, SC89820-000, Brazil
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
... ric acid, bile, and other factors (Pandey et al., 2021). This technology embeds probiotics in enteric-soluble wall material, which enhances their resistance to the adverse environment of the digestive tract. As a result, the probiotics can reach the intestine be released at the appropriate position, and exert their probiotic role (Cook et al., 2012). Dos Santos Wanderley et al. (2023 examined the in vitro application of saccharomyces microencapsulation in ruminant diets. Their investigation indicated that microencapsulated yeast had higher degradation potential and nutrient release than nonmicroencapsulated yeasts. In another study (Xiaofeng Chang et al., 2021), Lactobacillus acidophilus NCFM was successfully delive ...
Compound probiotics microcapsules improve milk yield and milk quality of dairy cows by regulating intestinal flora
Article
Full-text available
  • Mar 2024
To address the issues of probiotic activity loss during storage and feeding, as well as the limited efficacy of single probiotics, a solution was devised by embedding a mixture of Bacillus coagulans SN‐8 (SN‐8) and Saccharomyces boulardii SN‐6 (SN‐6) in a gel. The initial step involved screening the probiotic microcapsules' preparation method and wall material. Using sodium alginate and β‐cyclodextrin as composite wall material and chitosan as the outer coating material allowed for an embedding rate of 82.11% in composite probiotic microcapsules prepared by the air atomization method. Next, in vitro, simulated digestion experiments were conducted to determine the number of viable bacteria and the release rate of the microcapsules. The results showed that compared to the free strain, the mixed probiotic microcapsules retained a survival rate of 67.5% after 3 h of simulated gastric juice exposure and 70.56% after 42 days of storage at 4°C. This demonstrated higher survival rates and storage stability. The prepared probiotic microcapsules were then administered to dairy cows. 16S rDNA gene sequencing showed that consumption of the microcapsules reduced the number of harmful bacteria, such as Paeniclostridium , in the intestinal tract of dairy cows while accelerating the growth of beneficial bacteria, such as Bifidobacterium . In particular, this resulted in a significant improvement in the lactation performance of the cows, with a 4.5% increase in milk fat content, a 92.5% increase in milk protein content, and a 3.5% increase in milk urea nitrogen content (6.75 mL/dL). In conclusion, probiotic microcapsules can effectively regulate intestinal flora, improving milk production, and quality in dairy cows.
View
Role of probiotics in ruminant nutrition as natural modulators of health and productivity of animals in tropical countries: an overview
Article
Full-text available
  • Apr 2022
  • TROP ANIM HEALTH PRO
Given the ever-growing population in the developing countries located in the tropics of Asia, Africa, South America, and the Caribbean, the demand for products of animal origin has increased. Probiotics have proven to be a substantial substitute for antibiotics used in the animal diet and thus gained popularity. Probiotics are live and non-pathogenic microbes commercially utilized as modulators of gut microflora, hence exerting advantageous effects on the health and productivity of animals in tropical countries. Probiotics are mainly derived from a few bacterial (Lactobacillus, Enterococcus, Streptococcus, Propionibacterium, and Prevotella bryantii) and yeast (Saccharomyces and Aspergillus) species. Numerous studies in tropical animals revealed that probiotic supplementation in a ruminant diet improves the growth of beneficial rumen microbes, thus enhancing nutrient intake and digestibility, milk production, and reproductive and feed efficiency, along with immunomodulation. Furthermore, probiotic applications have proven to minimize adverse environmental consequences, including reduced methane emissions from ruminants’ anaerobic fermentation of tropical feedstuffs. However, obtained results were inconsistent due to sources of probiotics, probiotic stability during storage and feeding, dose, feeding frequency, and animal factors including age, health, and nutritional status of the host. Furthermore, the mechanism of action of probiotics by which they exhibit beneficial effects is still not clear. Thus, more definitive research is needed to select the most effective strains of probiotics and their cost–benefit analysis. In this review article, we have briefly explained the impact of feeding probiotics on nutrient intake, digestibility, reproduction, growth efficiency, productivity, and health status of tropical ruminant animals.
View
Metabolic engineering of the oleaginous yeast Yarrowia lipolytica PO1f for production of erythritol from glycerol
Article
Full-text available
  • Sep 2021
  • BIOTECHNOL BIOFUELS
Background Sugar alcohols are widely used as low-calorie sweeteners in the food and pharmaceutical industries. They can also be transformed into platform chemicals. Yarrowia lipolytica , an oleaginous yeast, is a promising host for producing many sugar alcohols. In this work, we tested whether heterologous expression of a recently identified sugar alcohol phosphatase (PYP) from Saccharomyces cerevisiae would increase sugar alcohol production in Y. lipolytica . Results Y. lipolytica was found natively to produce erythritol, mannitol, and arabitol during growth on glucose, fructose, mannose, and glycerol. Osmotic stress is known to increase sugar alcohol production, and was found to significantly increase erythritol production during growth on glycerol. To better understand erythritol production from glycerol, since it was the most promising sugar alcohol, we measured the expression of key genes and intracellular metabolites. Osmotic stress increased the expression of several key genes in the glycerol catabolic pathway and the pentose phosphate pathway. Analysis of intracellular metabolites revealed that amino acids, sugar alcohols, and polyamines are produced at higher levels in response to osmotic stress. Heterologous overexpression of the sugar alcohol phosphatase increased erythritol production and glycerol utilization in Y. lipolytica . We further increased erythritol production by increasing the expression of native glycerol kinase (GK), and transketolase (TKL). This strain was able to produce 27.5 ± 0.7 g/L erythritol from glycerol during batch growth and 58.8 ± 1.68 g/L erythritol during fed-batch growth in shake-flasks experiments. In addition, the glycerol utilization was increased by 2.5-fold. We were also able to demonstrate that this strain efficiently produces erythritol from crude glycerol, a major byproduct of the biodiesel production. Conclusions We demonstrated the application of a promising enzyme for increasing erythritol production in Y. lipolytica . We were further able to boost production by combining the expression of this enzyme with other approaches known to increase erythritol production in Y. lipolytica . This suggest that this new enzyme provides an orthogonal route for boosting production and can be stacked with existing designs known to increase sugar alcohol production in yeast such as Y. lipolytica . Collectively, this work establishes a new route for increasing sugar alcohol production and further develops Y. lipolytica as a promising host for erythritol production from cheap substrates such as glycerol.
View
Effect of probiotic lactobacilli supplementation on growth parameters, blood profile, productive performance, and fecal microbiology in feedlot cattle
Article
  • Feb 2023
  • RES VET SCI
Lactic acid bacteria (LAB) selected on the basis of probiotic characteristics were administered to beef feedlot catlle and the effect on body condition/growth and nutritional-metabolic status as well as on E. coli O157:H7 fecal shedding, were investigated. A feeding trials involving 126 steers were used to evaluate the effects of Lactobacillus acidophilus CRL2074, Limosilactobacillus fermentum CRL2085 and Limosilactobacillus mucosae CRL2069 and their combinations (5 different probiotic groups and control) when 107-108 CFU/animal of each probiotic group were in-feed supplemented. Cattle were fed a high energy corn-based diet (16 to 88%) and samples from each animal were taken at 0, 40, 104 and 163 days. In general, animals body condition and sensorium state showed optimal muscle-skeletal development and behavioral adaption to confinement; no nasal/eye discharges and diarrheic feces were observed. The nutritional performance of the steers revealed a steady increase of biometric parameters and weight. Animals supplied with L. mucosae CRL2069 for 104 days reached the maximum mean live weight (343.2 kg), whereas the greatest weight daily gain (1.27 ± 0.16 Kg/day) was obtained when CRL2069 and its combination with L. fermentum CRL2085 (1.26 ± 0.11 kg/day) were administered during the complete fattening cycle. With several exceptions, bovine cattle blood and serum parameters showed values within referential ranges. As a preharvest strategy to reduce Escherichia coli O157:H7 in cattle feces, CRL2085 administered during 40 days decreased pathogen shedding with a reduction of 43% during the feeding period. L. fermentum CRL2085 and L. mucosae CRL2069 show promise for feedlot cattle feeding supplementation to improve metabolic-nutritional status, overall productive performance and to reduce E. coli O157:H7 shedding, thus decreasing contamination chances of meat food products.
View
Soybean protein isolate treated with transglutaminase (TGase) enhances the heat tolerance of selected lactic acid bacteria strains to spray drying
Article
  • Oct 2022
  • FOOD CHEM
TGase treated soy protein isolate (SPI) was confirmed to improve the survival of Lb. bulgaricus CICC 6047 subjected to spray drying. The viability of this strain increased from 70.69% to 86.01% due to elevating thermal stability. The smoother bacterial cell surface was observed by TGase treated SPI. TGase treatment changed the secondary conformation of SPI, having α-helical structure increased. Chemical bonds (hydrogen bond, C-N bond, N-H bond) proved the enzyme-modified SPI to have enhanced resistance to heat. The addition of TGase treated SPI allowed other LAB strains to tolerate the spray drying better. Their survival percentage after spray drying was 83.44% for Le. mesenteroides CICC 6055, 80.64% for Lb. acidophilus NCFM, 87.79% for B. animalis BI-03 and 87.02% for Lb. plantarum CICC 6009, respectively. The good survival of the selected LAB strains from TGase treated SPI powders was observed during storage of 8 weeks at 4°C.
View
Enhanced survival of fluidized bed-dried microencapsulated Saccharomyces cerevisiae cells in the presence of Hongqu rice distiller's grain peptides
Article
  • Apr 2022
  • LWT-FOOD SCI TECHNOL
Saccharomyces cerevisiae (strain JH301) cells were micro-encapsulated in microcrystalline cellulose and Hongqu rice distiller's grain peptides (HRP) and subjected to fluidized bed drying. The effect of HRP on the viability of yeast in response to thermal drying was investigated. The moisture-viability profiles showed that HRP reduced the drying rate of yeast cells and increased viability in a concentration-dependent manner. Scanning electron microscopy revealed that HRP protected the morphology of the yeast cells, which presented a normal smooth structure and compact shape after thermal drying. The results of low-field ¹H nuclear magnetic resonance and infrared spectroscopy revealed that HRP controls the binding water of yeast powder as opposed to interacting with the membrane surface proteins, further reducing the damage to phospholipids in the cell membrane during drying. Flow cytometry and confocal laser scanning microscopy confirmed that 3% HRP could reduce the proportion of membrane-damaged cells by nearly 19%. After drying, the proportion of unsaturated fatty acids in the cell membrane of JH301-3% HRP was 10.36% higher than that of JH301 in the absence of HRP, achieving increased membrane integrity and higher ATPase activity after rehydration.
View
Effects of Saccharomyces cerevisiae var. boulardii on growth, incidence of diarrhea, serum immunoglobulins, and rectal microbiota of suckling dairy calves
Article
  • Feb 2022
  • LIVEST SCI
The present study was carried out to elucidate the growth, incidence of diarrhea, serum immunoglobulins, and rectal microbiota of dairy calves as they were supplemented with live Saccharomyces cerevisiae var. boulardii (S. cerevisiae var. boulardii). A total of 48 newborn Holstein dairy calves were selected as experimental animal. The calves were divided into four experimental groups based on the concentration of S. cerevisiae var. boulardii as feed supplement. The rate of feed supplement for the four groups were 0, 1, 3, and 5 g/day/calf of live S. cerevisiae var. boulardii, respectively. All animals of 0 were considered as control group. The study was continued for 42 days. Samples of blood and rectal contents were collected on d 21 and 42. The results showed that supplementation of S. cerevisiae var. boulardii improved the body weight, average daily gain, total feed intake, and feed conversion ratio during the first 21 d of life (P<0.05) of the experimental calves. Serum concentration of IgA, IgM, and IgG increased by 3 g/day and 5 g/day of treatments during the first 21 d of life (P<0.05). Moreover, supplementation with S. cerevisiae var. boulardii increased fecal score (3 and 5 g/day groups) and reduced the incidence of diarrhea (1, 3, and 5 g/day groups) during the first 21 d after birth (P<0.05). Both on d 21 and 42 of life, rectal samples were predominated by Firmicutes, Bacteroidetes, Fusobacteria, Proteobacteria, and Actinobacteria. Additionally, dietary supplementation with S. cerevisiae var. boulardii increased the relative abundance of genera Lactobacillus, Paracoccus, Blautia, and Eubacterium_oxidoreducens_group. However, compared to control, the relative abundance of Fusobacterium on d 21, was reduced (P<0.05). These findings indicated S. cerevisiae var. boulardii supplementation reduced the incidence of diarrhea, while improving serum immunoglobulins concentrations and modulating rectal microbiota in young dairy calves. Furthermore, we recommend that 3 g/day/calf S. cerevisiae var. boulardii supplemented feed be administered to dairy calves during their first 21 d of life.
View
Association between yeast product feeding and milk production of lactating dairy cows: Multilevel meta-analysis and meta-regression
Article
  • Feb 2022
  • ANIM FEED SCI TECH
To provide an overview of the association between commercially available yeast product (YP) supplementation and milk yield (MY) in lactating dairy cows, multilevel meta-analyses were performed on 99 trials from 49 peer-reviewed studies. Associations were evaluated using random effects models to examine the standardized mean difference (SMD) between the YP and control treatments. Associations were also quantified using the weighted mean difference (WMD). YP supplementation was associated with increased milk yield (+ 0.69 kg/d), milk fat content (+ 0.06%), milk fat yield (+ 0.04 kg/d) and milk protein yield (+ 0.02 kg/d). A positive association between YP and MY was not observed in primiparous cows during mid- and late lactation or in cows fed a high neutral detergent fiber diet. The improvement of MY in cases of YP supplementation was higher if YP supplementation started before calving (+ 0.79 kg/d) compared to after calving. The multivariate analyses showed that YP supplementation was associated with an increase in MY but not with an increase in dry matter intake (DMI). This provides strong evidence that greater milk production observed in supplemented cows is not a result of increased DMI.
View
Microencapsulation of probiotic starter culture in protein–carbohydrate carriers using spray and freeze-drying processes: Implementation in whey-based beverages
Article
  • Jan 2022
  • J FOOD ENG
The production of functional beverages has gained more attention because of growing consumers' interest in personal health. The process of carrier production and characterization of a novel type of protein-carbohydrate carriers for functional beverages with a focus on cell protection during fermentation, gastrointestinal, and storage conditions are shown in this study. Probiotic starter culture was microencapsulated using spray and freeze-drying techniques and then implemented in whey-based beverages. Encapsulation materials used are whey protein concentrate, whey, and sodium alginate. The encapsulation efficiency results were over 76.8% and 81.4%, after spray and freeze-drying respectively, depending on the carrier formulation. The culture in freeze-dried carriers reached a higher survival rate (>7.5 log10(CFUg⁻¹)) than one's encapsulated by spray-drying technique during storage. Microencapsulation of the culture significantly improved cell viability in gastrointestinal and storage conditions compared with free cells. Usage of both encapsulation techniques showed potential regarding cell preservation in production and storage.
View
Characteristic properties of spray-drying Bifidobacterium adolescentis microcapsules with biosurfactant
Article
  • Jan 2022
  • J BIOSCI BIOENG
The surfactants used for emulsion is one of the best techniques for microencapsulation of lactic acid bacteria (LAB) since it is economical. The biosurfactants have many advantages such as lower toxicity, higher biodegradability. In this study, microcapsules were prepared via spray drying using Bifidobacterium adolescentis species cultured in soy milk extract with biosurfactant prepared using Alcaligenes piechaudii CC-ESB2 to improve their powder properties. The soy milk was used to increase the health benefits instead of the milk. The optimum bacterial strain viability, water activity, and moisture content of the microcapsules were achieved at a spray dryer inlet/outlet temperature of 120/60°C. The composition of the carrier affects the particle size of the microcapsules. Using 90% maltodextrin (MD), 5% isomalto-oligosaccharide syrup (IMOS) and 5% biosurfactant as a carrier increased the viability of the LAB. Scanning electron microscope observations showed that the LAB microcapsules were able to effectively retain their completeness. Furthermore, microcapsules added with a biosurfactant prepared using A. piechaudii CC-ESB2 displayed significantly better flow properties than those without the surfactant and biosurfactant, which indicates that the biosurfactant assists in enhancing the powder properties of the microcapsules. It also has sufficient biological activity as a LAB product because the probiotics exceed 10⁶ CFU/mL The spray-dried abandoned supernatant with biosurfactant exhibited superior bacteriostasis, which suggests that the supernatant of B. adolescentis during microencapsulation not only retains its bacteriostatic effect under high spray drying temperatures, but also provides additional antibacterial effects for the microcapsules.
View
Microencapsulation with spray-chilling as an innovative strategy for probiotic low sodium requeijão cremoso processed cheese processing
Article
  • Dec 2021
The effect of sodium reduction and Lactobacillus acidophilus addition (free or microencapsulated) on the quality parameters of requeijão cremoso processed cheese was assessed for 90 days at 5 °C. Three formulations were prepared: (CONT) regular salt content and curd fusion (90 °C/2 min), (FREE) reduced salt content + free probiotic culture added after curd fusion (90 °C/2 min), and (MICRO) reduced salt content + microencapsulated probiotic culture added before curd fusion (70 °C/5 min). FREE and MICRO formulations showed increased monounsaturated and polyunsaturated fatty acid content, improved health indices (decreases in thrombogenic index and increases in desirable fatty acid index), and altered rheological properties (higher consistency index, apparent viscosity, elastic properties, and gel strength). In addition, the MICRO formulation showed higher texture sensory acceptance and probiotic counts higher than 6 log CFU/g during storage and simulated gastrointestinal conditions. Overall, microencapsulation of probiotics with spray chilling represents an innovative solution in requeijao cremoso processed cheese.
View